CN109913553A - Biomarker for diagnosing and treating breast cancer - Google Patents
Biomarker for diagnosing and treating breast cancer Download PDFInfo
- Publication number
- CN109913553A CN109913553A CN201910298609.4A CN201910298609A CN109913553A CN 109913553 A CN109913553 A CN 109913553A CN 201910298609 A CN201910298609 A CN 201910298609A CN 109913553 A CN109913553 A CN 109913553A
- Authority
- CN
- China
- Prior art keywords
- linc02237
- breast cancer
- product
- expression
- biomarker
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Pathology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Immunology (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Hospice & Palliative Care (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses the biomarker for diagnosing and treating breast cancer, the biomarker is LINC02237.The invention discloses a kind of product of Diagnosis of Breast cancer and detect application of the reagent of LINC02237 in the product for preparing Diagnosis of Breast cancer；The present invention discloses application of the LINC02237 in the computation model of building prediction breast cancer.
Description
Technical field
The invention belongs to biomedicine field, it is related to the biomarker for diagnosing and treating breast cancer, the biology
Marker is LINC02237.
Background technique
Tumour is the normal cell of structure function in body body, under the synergistic effect of inside and outside tumorigenesis factor, caused base
Because of horizontal abnormality and dysfunction, the hyperplasia of local anomaly and the neoformation that is formed.Common tumorigenesis substance includes chemistry
Tumorigen, virus and physics tumorigen.Newborn tumor tissues are normal with its source in cellular morphology and institutional framework
Tissue has differences, which is known as the atypia of tumor tissues.Malignant tumour, the cancer being just known as have infiltration
The characteristics of with transfer, and without complete coating, operation is not easy thoroughly to cut off.It is largely sought in addition, it can also consume body
Feeding substance is quickly bred, to the very harmful of body.For malignant tumour, our principle is early discovery, early to diagnose,
Early treatment.For finding later patient, the killer opportunity for the treatment of, the treatment pair such as radiotherapy, chemotherapy, targeted drug are often missed
It is very limited in the curative effect of patients with terminal, so that the survival rate sharp fall of patients with terminal.
Breast cancer is a kind of very common women's diseases, its classification includes many clinicopathologic features, such as tumour reality
Body size, whether occur lymphatic metastasis, histological grade and patient age distribution etc.；Patient makes during diagnosing and treating
With many biomarkers, mainly there are estrogen receptor (estrogen receptor ER), EGF-R ELISA
(human epidermal growth factor receptor 2, HER2), progesterone receptor (progesterone
Receptor, PR) etc..With people's deepening continuously to the research of breast cancer, breast cancer is summarized as 5 by newest genotyping result
Kind hypotype: HER2 overexpression type (ER-,PR-And HER2+), luminal A type (ER+Or PR+And HER2-), luminal Type B (ER+Or PR+And HER2+), substrate sample (basal-like) type (ER-,PR-And HER2-) and normal tissue (normal-like) type cream
Gland cancer (Mackay A, Weigelt B, Grigoriadis A, et al.Microarray-based class discovery
for molecular classification of breast cancer:analysis of interobserver
agreement[J].J Natl Cancer Inst,2011,103(8):662-673.).New parting is more conducive to for not
The clinical treatment of specificity is carried out with the patient with breast cancer of hypotype.
Long-chain non-coding RNA is that a kind of length is greater than 200 nucleotide and does not have the RNA of protein translation function,
Wide expression in transcriptional control.Currently, lncRNA is classified largely into five classes: antisense lncRNA, introne lncRNA, lincRNA,
Promoter correlation lncRNA lncRNA related to UTR.LncRNA has Space-time speciality, in different tissues, expression pattern row
It is also different.Relative to microRNA, the length of lncRNA is longer, has the similar structure of mRNA.LncRNA can be with
MicroRNA, mRNA and protein binding play important regulating and controlling effect in cell.At present, lncRNA and genetic transcription,
The biological activities such as epigenetic regulation, protein coding gene, chromatin organization are related.Meanwhile in RNA montage, x chromosome inactivation
Etc. in molecular mechanisms, lncRNAs also plays important function.LncRNA length is close with mRNA, has mRNA structure feature, can
To be combined with transcription factor, microRNA etc..Therefore, lncRNA plays the expression of protein coding gene and other non-coding RNAs
Compelling regulating and controlling effect is arrived.In addition to base sequence, lncRNA can also and protein binding, regulatory protein activity or exercise
Other functions.
Although the biological function of most of lncRNAs not yet determines, indicated there are many research work, cancer patient
LncRNA expression will appear exception in histocyte.In the research such as breast cancer, colorectal cancer, lung cancer, breast cancer, it was found that multiple
LncRNA is related to cancerous lesion.Unconventionality expression based on some lncRNAs is it is verified that, use closely related with cancer
LncRNAs also becomes a new research direction as cancer diagnosis and the marker of prognosis.
Summary of the invention
In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide a kind of relevant to breast cancer occurrence and development
The application of biomarker and the marker in breast cancer diagnosis and treatment.
To achieve the goals above, the present invention adopts the following technical scheme:
The present invention provides application of the reagent of detection LINC02237 in the product for preparing Diagnosis of Breast cancer.
Further, the breast cancer is Luminal Type B breast cancer.
Further, the product includes flat by RT-PCR, real-time quantitative PCR, in situ hybridization, chip or high-flux sequence
Platform detects the reagent of the expression of LINC02237.
Further, the reagent is selected from: the probe of specific recognition LINC02237；Or specific amplification LINC02237
Primer.
Further, the primer sequence of the specific amplification LINC02237 is as shown in NO.3~4 SEQ ID.
The present invention provides a kind of product of Diagnosis of Breast cancer, the product includes detection LINC02237 expression
Reagent.
Further, the product includes chip, kit, nucleic acid film item.
Further, the chip includes the oligonucleotide probe of specific recognition LINC02237；The kit includes spy
The primer of specific amplification LINC02237 or the oligonucleotide probe of specific recognition LINC02237；The nucleic acid film item includes
The oligonucleotide probe of specific recognition LINC02237.
Further, the primer sequence of specific amplification LINC02237 is as shown in NO.3~4 SEQ ID.
The present invention provides application of the LINC02237 in the computation model of building prediction breast cancer.
Further, the breast cancer is Luminal Type B breast cancer.
The present invention provides application of the LINC02237 in the pharmaceutical composition of preparation treatment breast cancer.
Further, the breast cancer is Luminal Type B breast cancer.
Further, described pharmaceutical composition includes the promotor of LINC02237.
Further, the promotor of the LINC02237 is the substance for increasing LINC02237 level.
Detailed description of the invention
Fig. 1 is the expression figure using QPCR detection LINC02237 gene in breast cancer tissue.
Specific embodiment
The present invention after extensive and in-depth study, by the method for high-flux sequence, detects in breast cancer sample
LncRNA has found lncRNA wherein with obvious differential expression, inquires into itself and cream in the expression of tumor tissues and normal tissue
Relationship between the generation of gland cancer, to find better approaches and methods for the diagnosis of breast cancer and targeted therapy.Pass through sieve
Choosing prompts LINC02237 to can be used as breast cancer present invention firstly discovers that LINC02237 is significantly raised in breast cancer tissue
Diagnosis marker and therapeutic targets.
Term " LINC02237 " is located on No. 8 chromosomes, gene I/D 105375706, including LINC02237 gene and
Its homologue, mutation and isoform.The term covers overall length, unprocessed LINC02237, and from processing in cell
Any type of LINC02237.The term covers the natural generation variant of LINC02237, and (such as splice variant or equipotential become
Body).The term covers such as LINC02237 gene, the gene order (NR_146282.1) of people LINC02237, and comes from and appoint
What its vertebrate origin, including mammal, such as Primate and rodent (such as mouse and rat)
LINC02237DNA。
It will be appreciated by those skilled in the art that the means of measurement gene expression are not importances of the invention.It can be
The expression of biomarker is detected on transcriptional level.The present invention can use any method known in the art measurement base
Because of expression.
As used in this article, term " biomarker ", which refers to, can detect in the sample and including such as LINC02237's
Index molecule or elements collection (such as predicting, diagnosis and/or prognostic indicator).Biomarker can be prediction biological marker
It object and serves as with specified disease or illness.Biomarker includes but is not limited to polynucleotides (such as DNA and/or RNA (example
Such as mRNA)), polynucleotide copies number changes (such as DNA copy number).
As used in this article, " amount " of biomarker or "horizontal" are the detectable levels in biological sample.This
It can be measured a bit by well known by persons skilled in the art and the methods disclosed herein.
Term " level of expression " or " expression " refer generally to the amount of biomarker in biological sample." expression " one
As refer to that information (such as gene coding and/or epigenetic information) is converted in cell the process of structure for existing and running.Cause
This, as used in this article, " expression ", which can refer to, is transcribed into polynucleotides, translates into polypeptide, or even polynucleotides and/or more
Peptide modifies (such as posttranslational modification of polypeptide).In specific embodiments of the present invention, " expression ", which refers to, is transcribed into multicore
Thuja acid.
" increased expression ", " increased expression ", " increased level ", " raised expression ", " raised expression water
It is flat " or " raised level " refer to relative to the control such as individual without disease or illness (such as cancer), internal contrast (example
Type of such as running one's home biomarker), or in patient group/group sample biomarker median expression level, it is a
The increased expression or increased level of biomarker in body.
" expression of reduction ", " expression of reduction ", " level of reduction ", " reduced expression ", " reduced expression water
It is flat " or " reduced level " refer to relative to control such as individual or internal contrast without disease or illness (such as cancer)
(such as type biomarker of running one's home), or in patient group/group sample biomarker median expression level,
The reduced expression or reduced level of biomarker in individual.In some embodiments, reduced expression is seldom
Expression or not.
In the present invention, LINC02237 has reduced expression in patient with breast cancer.
Chip, kit, nucleic acid film item
The present invention provides the product of the expression of LINC02237 gene in detection, the product includes (but unlimited
In) preparation, chip or kit.Wherein chip includes: solid phase carrier；And orderly it is fixed on the few core on the solid phase carrier
Thuja acid probe, the oligonucleotide probe some or all of specifically correspond to shown in LINC02237 sequence.
The solid phase carrier includes inorganic carrier and organic carrier, the inorganic carrier include but is not limited to have silicon carrier,
Glass carrier, ceramic monolith etc.；The organic carrier includes polypropylene film, nylon membrane etc..
As used herein, " oligonucleotides " refers generally to short, and single-stranded polynucleotides are less than about in length
250 nucleotide, but it's not necessary.Oligonucleotides can be synthesis.Term " oligonucleotides " and " polynucleotides " are simultaneously
It is not mutually exclusive.Description above for polynucleotides is same and is applicable to oligonucleotides completely.
Term " probe " refers to can be with the molecule in conjunction with the particular sequence of another molecule or subsequence or other parts.Unless another
It points out, term " probe " is often referred to match and another polynucleotides (often referred to as " target polynucleotide ") by complementary base
In conjunction with polynucleotide probes.Lack sufficient sequence complementarity according to the stringency of hybridization conditions, probe energy and with the probe
Target polynucleotide combines.Probe can make direct or indirect label, and range includes primer.Crossing system, including, but it is unlimited
In: solution phase, solid phase, mixed phase or in situ hybridization measuring method.
These probes have the base sequence with the specific base sequence complementary of target gene.Here, so-called " complementation ",
As long as hybridization, can not be complete complementary.These polynucleotides have usually relative to the specific base sequence
80% or more, preferably 90% or more, more preferable 95% or more, particularly preferred 100% homology.These probes can be DNA,
It is also possible to RNA, furthermore it is possible to pass through PNA (Polyamide nucleic in part of it or whole nucleotides
Acid, peptide nucleic acid), LNA (registered trademark, locked nucleic acid, Bridged Nucleic Acid, Cross-linked core
Acid), ENA (registered trademark, 2 '-O, 4 '-C-Ethylene-bridged nucleic acids), GNA (Glycerol
Nucleic acid, glycerol nucleic acid), the artificial replacement nucleic acid such as TNA (Threose nucleic acid, threose nucleic acid) obtains
Polynucleotides.
Kit of the invention includes the reagent for detecting LINC02237 gene, one or more substances selected from the group below: is held
Device, operation instructions, positive control, negative control object, buffer, auxiliary agent or solvent.
In kit of the invention can also have kit operation instructions, be described how using kit into
Row detection, and how tumor development to be judged using testing result, therapeutic scheme is selected.
The component of kit can pack in the form of aqueous medium or in the form of freeze-drying.Container appropriate in kit
It include typically at least a kind of bottle, test tube, flask, PET bottle, syringe or other containers, wherein a kind of component can be placed, and
And preferably, suitably equal part can be carried out.There are more than one group timesharing in kit, generally also will include in kit
Second, third or other additional containers, wherein being positioned separately additional component.However, the component of various combination can be wrapped
It is contained in a bottle.Kit of the invention will include generally also a kind of container for being used to accommodate reactant, seal to be used for
Commercial distribution.This container may include the plastic containers of injection molding or blowing mould, wherein can retain required bottle.
Nucleic acid film item of the invention includes that substrate and the oligonucleotides for LINC02237 being fixed in the substrate are visited
Needle；The substrate can be any substrate suitable for immobilized oligonucleotide probe, such as nylon membrane, nitrocellulose filter, poly- third
Alkene film, sheet glass, silica gel chip, miniature magnetic bead etc..
Computation model
The present invention provides application of the LINC02237 in the computation model of preparation prediction breast cancer.As knack
What personnel can be appreciated that, the measurement of two or more markers can be used to improve the diagnosis problem in investigation.Biochemistry mark
Will object can measure individually, or in one embodiment of the invention, they can be measured simultaneously, for example, using chip or
Array technique based on pearl.Then the independent concentration for interpreting biomarker, such as using individual retentions of every kind of marker, or
Their combinations of person are interpreted.
In the present invention, it can be implemented in various ways marker levels and certain possibility or risk association and realize
The step of getting up.Preferably, the mathematically measurement concentration of combination gene and one or more other markers, and by combined value
It associates with basic diagnosis problem.It can be by any suitable prior art mathematical method by the measurement group of marker levels
It closes.
Promotor
The promotor of the LINC02237 refer to it is any can be improved LINC02237 gene or expression product stability, on
The expression of LINC02237 is adjusted, the effective acting time of lncRNALINC02237 is increased or promotes the transcription of LINC02237 gene
Substance, these substances are used equally for the present invention, the substance useful as the expression for up-regulation LINC02237 gene, thus
It can be used for preventing or treating breast cancer.
The present invention is described in further detail with reference to the accompanying drawings and examples.Following embodiment is merely to illustrate this
It invents rather than limits the scope of the invention.Test method without specific conditions in embodiment, usually according to conventional strip
Part, or according to the normal condition proposed by manufacturer.
Embodiment 1 screens gene marker relevant to breast cancer
1, sample collection
The cancerous tissue and corresponding normal tissue sample for collecting 4 Luminal Type B breast cancer respectively are (apart from tumour side
5 centimeters of edge), high-flux sequence is carried out, all patients are preoperative not to carry out chemotherapy, radiotherapy and endocrine therapy, and all patients are equal
Informed consent, the acquirement of above-mentioned all samples pass through the agreement of the committee, organizational ethics, and patient information is as shown in table 1.
1 sample information of table
2, the preparation and quality analysis of RNA sample
Total tissue RNA is extracted using TRIZOL method
1) it is shredded with scissors tissue, 1ml Trizol is added, shakes 1min on oscillator；Room temperature 10min makes core egg
Lean type is decomposed completely.
2) 200 μ l chloroforms (chloroform) are added, cover tightly pipe lid, acutely shake 15s, room temperature stands 10min.
3) 4 DEG C, 11000rpm is centrifuged 15min.
4) water sample layer is transferred in a new centrifuge tube, 500 μ l isopropanols is added；After being mixed by inversion, room temperature is stood
10min。
5) 4 DEG C, 11000rpm is centrifuged 15min.
6) liquid is carefully siphoned away with rifle, stays and is deposited in tube bottom, the ethyl alcohol of 1ml 75% is added, shakes 5s on the oscillator,
Washing precipitating is primary.
7) 4 DEG C, 8000rpm is centrifuged 5min.
8) supernatant is carefully removed, drying precipitated 10min, suitable water dissolution precipitating 10min is added.
9) RNA concentration is detected, identifies the yield and purity of RNA.
3, the building and sequencing of cDNA library
1) total serum IgE DNaseI digests: using DNA fragmentation present in DNase I digestion Total RNA sample, magnetic bead is pure
Change recycling reaction product, is finally dissolved in DEPC water；
2) rRNA: the good Total RNA sample of cancellationization is removed, is removed using the Ribo-Zero kit of Epicentre
RRNA carries out Agilent 2100 after removal and detects, verifies rRNA removal effect；
3) RNA is interrupted: taking previous step sample, addition interrupts Buffer, is placed in progress heat in PCR instrument and interrupts, interrupts
140-160nt；
4) synthesis of one chain of reverse transcription: appropriate primer being added into the sample after interrupting, after mixing well
Thermomixer thermophilic reacts certain time, is allowed to open secondary structure and in conjunction with primer, adds the chain prepared in advance
Synthetic reaction system Mix synthesizes a chain cDNA by corresponding program in PCR instrument；
5) synthesis of two chain of reverse transcription: preparing two chain synthesis reaction systems, one timing of thermophilic reaction on Thermomixer
Between, synthesis has the two chain cDNA of dUTP, and reaction product carries out purification and recovery with magnetic bead；
6) end is repaired: being prepared end and is repaired reaction system, thermophilic reacts certain time in Thermomixer, in enzyme
Under the action of, the cohesive end for the cDNA double-strand that reverse transcription obtains is repaired, end reparation product is purified with magnetic bead
Recycling, is finally dissolved in EB Solution for sample；
7) 3 ' end cDNA adds " A ": it prepares and adds " A " reaction system, thermophilic reacts certain time in Thermomixer,
Under the action of enzyme, 3 ' ends for the product cDNA for repairing end are plus A base；
8) connection of 5 ' adapter of cDNA: preparing connector coupled reaction system, the thermophilic reaction one in Thermomixer
It fixes time, under the action of enzyme, connect connector with A base, product carries out purification and recovery with magnetic bead；
9) UNG digests bis- chain of cDNA: UNG digestion reaction system is prepared, two chains in double-stranded DNA are fallen by UNG enzymic digestion,
And purification and recovery is carried out to product with magnetic bead；
10) PCR reaction and product recycling: PCR reaction system is prepared, PCR response procedures appropriate is selected, upper step is obtained
To product expanded, to PCR product carry out magnetic beads for purifying recycling, recovery product is dissolved in EB solution, labelled.
11) Library Quality detects: using Agilent 2100Bioanalyzer and ABI StepOnePlus Real-
Time PCR System detects Library Quality；
12) machine is sequenced on: detecting qualified library, NaOH denaturation is added at single-stranded, it is anticipated that upper machine data volume, dilution
To certain upper machine concentration.Library after denaturation dilution is added in FlowCell, is hybridized with the connector on FlowCell,
Bridge-type PCR amplification is completed on cBot, is finally sequenced using IlluminaHiseq x-ten platform.
4, bioinformatic analysis
1) with cutadapt to the 5 ' of reads and 3 ' Duan Jinhang trim, trim falls the base of quality < 20, and it is big to delete N
In 10% reads；
2) hisat2 is compared onto reference genome.With reference to genome from Ensembl database, genome version
GRCh38, gene annotation information are Ensemble 92；
3) stringtie quantifies the expression quantity and normalization output of lncRNA；
4) edgeR packet compares control group with the differential expression of disease group lncRNA, the screening criteria of variances movement lncRNA
It is | log2FC |>1 and pvalue<0.05.
5, result
Sequencing data is as shown in table 2, and bioinformatic analysis discovery, LINC02237 is expressed significantly in patient with breast cancer
It lowers, LINC02237 is prompted to can be used as the early diagnosis that possible detection target is applied to breast cancer.
2 sequencing data of table
The differential expression of 2 QPCR sequence verification LINC02237 gene of embodiment
1, according to 25 luminal Type B breast cancer patients tissue samples of the collection mode collection of embodiment 1 and just
Normal tissue samples carry out large sample QPCR verifying to LINC02237 gene differential expression.
2, RNA is extracted
Tissue RNA is extracted using Trizol method, specific steps are referring to embodiment 1.
3, it reverse transcription: is operated using the reverse transcription reagent box (Takara code:DRR047A) of TAKARA company.
1) genomic DNA is removed
5 × gDNA Eraser B μ ffer, 2.0 μ l, gDNA Eraser1.0 μ l is added in test tube, 1 μ g of total serum IgE adds
Rnase Free ddH2O makes total volume to 10 μ l, 42 DEG C of heating 2min in water-bath.
2) reverse transcription reaction
It will24.0 μ l of Buffer,RT Enzyme Mix I1.0 μ l, RT
Primer Mix1.0 μ l, RNase Free ddH24.0 μ l of O, which is added in above-mentioned test tube, is mixed together totally 20 μ l, 37 in water-bath
DEG C 15min, 85 DEG C of 5s.
4, QPCR is expanded
1) design of primers
According to the gene order design primer of LINC02237 and GADPH, specific primer sequence is as follows:
GAPDH gene:
Forward primer is 5 '-AATCCCATCACCATCTTCCAG-3 ' (SEQ ID NO.1)；
Reverse primer is 5 '-GAGCCCCAGCCTTCTCCAT-3 ' (SEQ ID NO.2).
LINC02237 gene:
Forward primer is 5 '-ACACAATTCTAACTCTCA-3 ' (SEQ ID NO.3)；
Reverse primer is 5 '-TTCCTCCATTTGAAGATA-3 ' (SEQ ID NO.4).
2) QPCR amplification is examined
WithPremix Ex TaqTMII (Takara Code:DRR081) kit configures PCR reaction system,
Thermal CyclerPCR amplification is carried out on Real Time System amplification instrument, confirms Real after reaction
The amplification curve and solubility curve of Time PCR, Δ Δ CT method carry out relative quantification.
Configure 25 μ l reaction systems:
Premix Ex TaqTM II (2 ×) 12.5 μ l, it is positive (anti-) to go out to each 1 μ l of primer, 2 μ l of DNA profiling
8.5 μ l of bacterium distilled water.
Reaction condition: 95 DEG C of 30s, (95 DEG C of 5s, 60 DEG C of 30s) × 40
5, result
QPCR result is as shown in Figure 1, compared with normal tissue, and LINC02237 is lowered in expression in breast, difference
It is consistent with high-flux sequence result with statistical significance (P < 0.05), prompt LINC02237 to can be used as biomarker application
In the diagnosing and treating of breast cancer.
The explanation of above-described embodiment is only intended to understand method and its core concept of the invention.It should be pointed out that for this
For the those of ordinary skill in field, without departing from the principle of the present invention, several improvement can also be carried out to the present invention
And modification, these improvement and modification will also be fallen into the protection scope of the claims in the present invention.
Sequence table
<110>People's Hospital of Deyang City
<120>it is used for the biomarker of diagnosing and treating breast cancer
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
aatcccatca ccatcttcca g 21
<210> 2
<211> 19
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
gagccccagc cttctccat 19
<210> 3
<211> 18
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
acacaattct aactctca 18
<210> 4
<211> 18
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
ttcctccatt tgaagata 18
Claims (10)
1. detecting application of the reagent of LINC02237 in the product for preparing Diagnosis of Breast cancer.
2. application according to claim 1, which is characterized in that the breast cancer is Luminal Type B breast cancer.
3. application according to claim 2, which is characterized in that the product include by RT-PCR, real-time quantitative PCR,
The reagent of the expression of in situ hybridization, chip or high-flux sequence detection of platform LINC02237.
4. application according to claim 1, which is characterized in that the reagent is selected from: the spy of specific recognition LINC02237
Needle；Or the primer of specific amplification LINC02237.
5. application according to claim 4, which is characterized in that the primer sequence of the specific amplification LINC02237 is such as
Shown in NO.3~4 SEQ ID.
6. a kind of product of Diagnosis of Breast cancer, which is characterized in that the product includes the examination for detecting LINC02237 expression
Agent.
7. product according to claim 6, which is characterized in that the product includes chip, kit, nucleic acid film item.
8. product according to claim 7, which is characterized in that the chip includes the widow of specific recognition LINC02237
Nucleotide probe；The kit includes the primer of specific amplification LINC02237 or the widow of specific recognition LINC02237
Nucleotide probe；The nucleic acid film item includes the oligonucleotide probe of specific recognition LINC02237.
9.LINC02237 the application in the computation model of building prediction breast cancer.
10.LINC02237 the application in the pharmaceutical composition of preparation treatment breast cancer.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910298609.4A CN109913553A (en) | 2019-04-15 | 2019-04-15 | Biomarker for diagnosing and treating breast cancer |
| CN201911075763.1A CN110714080B (en) | 2019-04-15 | 2019-11-06 | Biomarkers for diagnosis and treatment of breast cancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910298609.4A CN109913553A (en) | 2019-04-15 | 2019-04-15 | Biomarker for diagnosing and treating breast cancer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109913553A true CN109913553A (en) | 2019-06-21 |
Family
ID=66969810
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910298609.4A Pending CN109913553A (en) | 2019-04-15 | 2019-04-15 | Biomarker for diagnosing and treating breast cancer |
| CN201911075763.1A Active CN110714080B (en) | 2019-04-15 | 2019-11-06 | Biomarkers for diagnosis and treatment of breast cancer |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911075763.1A Active CN110714080B (en) | 2019-04-15 | 2019-11-06 | Biomarkers for diagnosis and treatment of breast cancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN109913553A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110714080A (en) * | 2019-04-15 | 2020-01-21 | 德阳市人民医院 | Biomarkers for diagnosis and treatment of breast cancer |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112921098B (en) * | 2021-04-27 | 2023-05-05 | 深圳市人民医院 | Marker for detecting triple negative breast cancer, detection reagent and application thereof |
| CN113817832B (en) * | 2021-10-22 | 2023-05-05 | 大理大学 | LincRNA marker for predicting prognosis of breast cancer, detection reagent and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106222242A (en) * | 2016-01-29 | 2016-12-14 | 复旦大学 | Long-chain non-coding RNA-APOC1P1-3 gene and for preparing the purposes in target spot mark |
| CN107937550A (en) * | 2018-01-08 | 2018-04-20 | 北京泱深生物信息技术有限公司 | A kind of and the relevant biomarker of breast cancer occurrence and development and its application |
| CN108034725A (en) * | 2018-01-08 | 2018-05-15 | 北京泱深生物信息技术有限公司 | Applications of the LINC02185 in breast cancer diagnosis and treatment |
| CN108949988A (en) * | 2018-08-03 | 2018-12-07 | 武汉大学 | A kind of application of long-chain non-coding RNA SNHG6 in breast cancer diagnosis or treatment |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146693B (en) * | 2013-02-26 | 2014-07-09 | 中南大学 | Long chain non-coding RNA (Ribonucleic Acid) gene and application method thereof |
| CN108085389B (en) * | 2017-12-29 | 2020-06-09 | 青岛泱深生物医药有限公司 | LncRNA related to breast cancer and application thereof |
| CN109913553A (en) * | 2019-04-15 | 2019-06-21 | 德阳市人民医院 | Biomarker for diagnosing and treating breast cancer |
-
2019
- 2019-04-15 CN CN201910298609.4A patent/CN109913553A/en active Pending
- 2019-11-06 CN CN201911075763.1A patent/CN110714080B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106222242A (en) * | 2016-01-29 | 2016-12-14 | 复旦大学 | Long-chain non-coding RNA-APOC1P1-3 gene and for preparing the purposes in target spot mark |
| CN107937550A (en) * | 2018-01-08 | 2018-04-20 | 北京泱深生物信息技术有限公司 | A kind of and the relevant biomarker of breast cancer occurrence and development and its application |
| CN108034725A (en) * | 2018-01-08 | 2018-05-15 | 北京泱深生物信息技术有限公司 | Applications of the LINC02185 in breast cancer diagnosis and treatment |
| CN108949988A (en) * | 2018-08-03 | 2018-12-07 | 武汉大学 | A kind of application of long-chain non-coding RNA SNHG6 in breast cancer diagnosis or treatment |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110714080A (en) * | 2019-04-15 | 2020-01-21 | 德阳市人民医院 | Biomarkers for diagnosis and treatment of breast cancer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110714080B (en) | 2020-09-15 |
| CN110714080A (en) | 2020-01-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109609650B (en) | Biomarkers for diagnosis and treatment of hepatocellular carcinoma | |
| WO2008002672A2 (en) | Targets for use in diagnosis, prognosis and therapy of cancer | |
| EP2800820A1 (en) | Methods and kits for detecting subjects at risk of having cancer | |
| CN109609648A (en) | LncRNA marker relevant to liver cancer and its detection primer and application | |
| CN109913553A (en) | Biomarker for diagnosing and treating breast cancer | |
| Kohashi et al. | Differential microRNA expression profiles between malignant rhabdoid tumor and epithelioid sarcoma: miR193a-5p is suggested to downregulate SMARCB1 mRNA expression | |
| CN110628913B (en) | lncRNA marker related to breast cancer | |
| CN106929577B (en) | lncRNA biomarker related to lung adenocarcinoma | |
| WO2011128820A2 (en) | Methods for the analysis of breast cancer disorders | |
| CN110257516A (en) | For developing molecular marker and the application of diagnosing gastric cancer product | |
| CN110157808A (en) | A kind of application of non-coding RNA relevant to larynx squamous carcinoma occurrence and development | |
| CN111893188B (en) | Application of biomarker LINC01356 in diagnosis and treatment of cervical cancer | |
| CN109825595A (en) | LncRNA marker relevant to breast cancer and its detection primer and application | |
| CN111979315A (en) | Application of annular TP63 as lung squamous carcinoma diagnosis or treatment target | |
| CN113637763A (en) | Application of miRNA biomarker in early diagnosis and treatment of melanoma | |
| CN109913552A (en) | A kind of esophageal squamous cell carcinoma diagnosis and treatment target spot and application | |
| CN110055333A (en) | Application of the RP11-116O18.1 as molecular marker in lung cancer | |
| CN110592227B (en) | Application of biomarker in breast cancer | |
| CN110205385A (en) | Molecular marker and its application for adenocarcinoma of lung diagnosis | |
| CN105821131B (en) | Osteosarcoma miRNA marker | |
| CN109628598A (en) | Application of the long-chain non-coding RNA in tumour | |
| CN110117656A (en) | A kind of biomarker relevant to sdenocarcinoma of stomach occurrence and development | |
| CN110066876A (en) | A kind of molecular marker for cancer diagnosis | |
| CN109609649A (en) | A kind of lncRNA for rectal adenocarcinoma diagnosis and treatment | |
| CN112553333B (en) | Application of miR-1207 and target gene thereof in detection of laryngeal squamous cell carcinoma |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190621 |